Fernandes Helen, Koneru Baburao, Fernandes Neil, Hameed Meera, Cohen Marion C, Raveche Elizabeth, Cohen Stanley
Department of Pathology and Laboratory Medicine, UMDNJ-New Jersey Medical School, Newark, New Jersey 07103, USA.
Transplantation. 2002 Jun 27;73(12):1886-91. doi: 10.1097/00007890-200206270-00006.
Cytokines such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-10 play significant roles in the inflammatory and immune responses that mediate allograft rejection. The presence of a G-->A polymorphism at position -308 in the promoter region of the TNF-alpha gene increased its transcription 6- to 7-fold. A similar polymorphism at position -1082 of the IL-10 promoter results in decreased production of IL-10 protein. In this study we have determined whether the single nucleotide polymorphisms in the promoter regions of the TNF-alpha and IL-10 genes can predict the outcome of the allograft in liver recipients.
DNA was extracted from whole blood of liver recipients. The genotype of the patients was determined by polymerase chain reaction using sequence-specific primers. The level of TNF-alpha and IL-10 protein was measured by ELISA after stimulation of peripheral blood mononuclear cells with concanavalin A.
There was significant correlation between acute cellular rejection and the presence of the -308A polymorphism (P<0.001), with 8 of 13 patients with the TNF-alpha polymorphism having evidence of acute rejection. Cell stimulation studies revealed that the level of TNF-alpha protein produced by patients with liver rejection was significantly higher than for patients without rejection (P=0.001). There were no strong associations between the presence of the IL-10 polymorphisms and rejection (P=0.71).
This study adds to the understanding of the role of cytokine polymorphisms in liver transplants. The data suggest that cytokine promoter polymorphisms may be a risk factor associated with allograft rejection in the liver.
细胞因子如肿瘤坏死因子（TNF）-α和白细胞介素（IL）-10在介导同种异体移植排斥反应的炎症和免疫反应中发挥重要作用。TNF-α基因启动子区域-308位点存在G→A多态性，可使其转录增加6至7倍。IL-10启动子-1082位点的类似多态性导致IL-10蛋白产生减少。在本研究中，我们确定了TNF-α和IL-10基因启动子区域的单核苷酸多态性是否可预测肝移植受者的移植结果。
从肝移植受者的全血中提取DNA。使用序列特异性引物通过聚合酶链反应确定患者的基因型。用伴刀豆球蛋白A刺激外周血单个核细胞后，通过酶联免疫吸附测定法测量TNF-α和IL-10蛋白水平。
急性细胞排斥反应与-308A多态性的存在之间存在显著相关性（P<0.001），13例具有TNF-α多态性的患者中有8例有急性排斥反应的证据。细胞刺激研究显示，肝移植排斥患者产生的TNF-α蛋白水平显著高于无排斥反应的患者（P=0.001）。IL-10多态性的存在与排斥反应之间没有强关联（P=0.71）。
本研究增进了对细胞因子多态性在肝移植中作用的理解。数据表明，细胞因子启动子多态性可能是与肝移植排斥相关的一个危险因素。
